Site Of Administration Research Articles

PL02.1.A EO2401, a novel microbiome-derived therapeutic vaccine for patients with recurrent glioblastoma: ROSALIE study

Abstract Background EO2401 (EO) was designed to activate existing commensal memory T-cells cross-reacting with tumor associated antigens (TAAs). EO includes microbial-derived, synthetically produced peptides corresponding to HLA-A2 restricted epitopes with molecular mimicry to three TAAs upregulated in glioblastoma (GB), IL13Rα2, BIRC5 and FOXM1, with the CD4 helper peptide UCP2 and the adjuvant Montanide. Pre-clinically EO generated strong immune responses and cross-reactive CD8 cells recognizing the targeted TAAs. Methods This ongoing Ph 1/2 trial (NCT04116658) investigates the safety and tolerability (primary) of EO (300 µg/peptide, SC Q2W X 4, then Q4W), EO with nivolumab (3 mg/kg Q2W; EN), and EN with bevacizumab (10 mg/kg Q2W; ENB) among four Cohorts (Cs) of pts with GB at first progression/recurrence after radiotherapy/temozolomide. Treatment was delivered until progression, or 24 months. After the Ph 1 of EO followed by EN (C1), C2 investigated EN without (C2a) or with (C2b) surgery while C3 investigated ENB (population as C2a). Results Among 40 treated pts (C1 n=3, C2a n=23, C2b n=3, C3 n=11), median age was 60 years, 53% were male, 40% had KPS 90-100%, 35% had O6-methylguanine DNA-methyltransferase promotor hypermethylated tumors, and 5% isocitrate dehydrogenase 1 mutated tumors. All evaluable pts demonstrated strong CD8 T-cell ELISPOT responses against the 3 vaccine peptides; response was shown with tetramer staining of specific CD8 in 24/25 investigated pts after in vitro stimulation and in 19/20 pts directly ex vivo. Cross-reactivity against targeted TAAs was confirmed in 20/21 pts. Majority of response were detected by week 4 after 1st dose and as early as 2 weeks in some pts. EO, EN, and ENB were well tolerated (max exposure EN 86 wks, ENB 47 wks) with EO associated toxicity limited to local administration site reactions (48%; all grade 1-2). The frequency and severity of nivolumab- or bevacizumab-associated AEs was consistent with historical single-agent profiles. With a median follow-up of 13.6 months, median progression-free survival (mPFS), and median survival for EN (C1+C2a+C2b) were 1.8 months (2 ongoing at 7.3, and 18.5 months), and 11.0 months (survival at 12 months 42%), respectively. With a median follow-up of 7.3 months (range, 3.0-10.5), pts on ENB (C3) have mPFS and survival at 6 months of 5.5 months (3 ongoing at 5.6, 7.3 and 9.1 months), and 82% (9/11 alive), respectively. ORR/DCR (ORR+SD) for EN and ENB were 10%/34% and 55%/82%, respectively. Conclusion EO2401 generated strong systemic immune responses and was well tolerated in combination with nivolumab ± bevacizumab. Preliminary ORR/DCR and mPFS for ENB, and survival for EN seem encouraging. Updated results from the current 40 patients and results from additional 35 patients who already started treatment with EN with the option for low-dose bevacizumab edema treatment at neurological symptoms will be presented.

The Mucoadhesive Nanoparticle-Based Delivery System in the Development of Mucosal Vaccines.

Mucosal tissue constitutes the largest interface between the body and the external environment, regulating the entry of pathogens, particles, and molecules. Mucosal immunization is the most effective way to trigger a protective mucosal immune response. However, the majority of the currently licensed vaccines are recommended to be administered by intramuscular injection, which has obvious shortcomings, such as high production costs, low patient compliance, and lack of mucosal immune response. Strategies for eliciting mucosal and systemic immune responses are being developed, including appropriate vaccine adjuvant, delivery system, and bacterial or viral vectors. Biodegradable mucoadhesive nanoparticles (NPs) are the most promising candidate for vaccine delivery systems due to their inherent immune adjuvant property and the ability to protect the antigen from degradation, sustain the release of loaded antigen, and increase the residence time of antigen at the administration site. The current review outlined the complex structure of mucosa, the mechanism of interaction between NPs and mucosa, factors affecting the mucoadhesion of NPs, and the application of the delivery system based on mucoadhesive NPs in the field of vaccines. Moreover, this review demonstrated that the biodegradable and mucoadhesive NP-based delivery system has the potential for mucosal administration of vaccines.

Quantifying inequities in COVID-19 vaccine distribution over time by social vulnerability, race and ethnicity, and location: A population-level analysis in St. Louis and Kansas City, Missouri.

Equity in vaccination coverage is a cornerstone for a successful public health response to COVID-19. To deepen understanding of the extent to which vaccination coverage compares with initial strategies for equitable vaccination, we explore primary vaccine series and booster rollout over time and by race/ethnicity, social vulnerability, and geography. We analyzed data from the Missouri Department of Health and Senior Services on all COVID-19 vaccinations administered across 7 counties in the St. Louis region and 4 counties in the Kansas City region. We compared rates of receiving the primary COVID-19 vaccine series and boosters relative to time, race/ethnicity, zip-code-level Social Vulnerability Index (SVI), vaccine location type, and COVID-19 disease burden. We adapted a well-established tool for measuring inequity-the Lorenz curve-to quantify inequities in COVID-19 vaccination relative to these key metrics. Between 15 December 2020 and 15 February 2022, 1,763,036 individuals completed the primary series and 872,324 received a booster. During early phases of the primary series rollout, Black and Hispanic individuals from high SVI zip codes were vaccinated at less than half the rate of White individuals from low SVI zip codes, but rates increased over time until they were higher than rates in White individuals after June 2021; Asian individuals maintained high levels of vaccination throughout. Increasing vaccination rates in Black and Hispanic communities corresponded with periods when more vaccinations were offered at small community-based sites such as pharmacies rather than larger health systems and mass vaccination sites. Using Lorenz curves, zip codes in the quartile with the lowest rates of primary series completion accounted for 19.3%, 18.1%, 10.8%, and 8.8% of vaccinations while representing 25% of the total population, cases, deaths, or population-level SVI, respectively. When tracking Gini coefficients, these disparities were greatest earlier during rollout, but improvements were slow and modest and vaccine disparities remained across all metrics even after 1 year. Patterns of disparities for boosters were similar but often of much greater magnitude during rollout in fall 2021. Study limitations include inherent limitations in the vaccine registry dataset such as missing and misclassified race/ethnicity and zip code variables and potential changes in zip code population sizes since census enumeration. Inequities in the initial COVID-19 vaccination and booster rollout in 2 large US metropolitan areas were apparent across racial/ethnic communities, across levels of social vulnerability, over time, and across types of vaccination administration sites. Disparities in receipt of the primary vaccine series attenuated over time during a period in which sites of vaccination administration diversified, but were recapitulated during booster rollout. These findings highlight how public health strategies from the outset must directly target these deeply embedded structural and systemic determinants of disparities and track equity metrics over time to avoid perpetuating inequities in healthcare access.

Botulinum toxin (BoNT) in the correction of mimic wrinkles - the most common complications

Introduction and purpose
 Botulinum toxin (BoNT) products are commonly used to treat muscle spasms such as cervical dystonia as well as to treat expression lines. Serotype A of botulinum toxin is the strongest and is used in aesthetic medicine procedures. It provides predictable results while having few side effects. Nevertheless, complications may occur, such as: ptosis of the upper eyelid, drooping eyebrows, ectropia, double vision, xerophthalmia, falling corners of the mouth, dysphagia, hoarseness, local swelling, erythema, bruising, pain at the injection site, asymmetry.
 The aim of the study is to discuss the most common complications after injection of the botulin toxin in the correction of mimic wrinkles.
 Description of the state of knowledge
 BoNT is a neurotoxin that acts on the neuromuscular junctions, inhibits the release of acetylcholine, consequently causing temporary chemical denervation. Muscle function begins to return approximately 3 months after injection and fully returns after 6 months. Comparison of clinical trials of botulinum toxin type A in aesthetic allowed for the quantification of the frequency of adverse effects:drooping eyelids (2.5%), drooping eyebrows (3.1%), disturbed eye sensation (3%) and lip asymmetry (6.9%). Complications after cosmetic botulinum toxin injections are rare, and those that do occur are usually mild and transient. These complications are technique dependent; the incidence decreases as the injection ability improves.
 Summary
 Side effects are rare, nevertheless, thorough knowledge of the patient's anatomy, medical history, possible complications related to the specific product and site of administration, and close monitoring are essential for safe, effective treatment and the achievement of satisfactory results.

The influence of different bioadhesive polymers on physicochemical properties of thermoresponsive emulgels containing Amazonian andiroba oil

Emulgels are amphiphilic semi-solid that can control the delivery of drugs showing different polarities. The use of a polymeric matrix in the hydrophilic portion leads to a higher thermodynamic stability system. Emulgels composed of poloxamer 407 (P407) and a bioadhesive polymer has shown improved contact time and adhesiveness on the site of administration, which can lead to an increase in the bioavailability of drugs. This work studied the effect of three different bioadhesive polymers, namely Carbopol 974P (C974P), polycarbophil (PCB) and poly(vinyl methyl ether-alt-maleic anhydride) (PMV/MA), on physicochemical characteristics of emulsion systems containing P407 and Amazonian andiroba oil. This oil is utilized as emollient, humectant and antioxidant. A factorial design 34 was utilized, and the physicochemical stability was investigated. The mechanical textural and rheological properties of the most stable formulations were evaluated for selection of potential emulsion systems. According to the texture profile and softness analyses, the selected formulations were mainly influenced by the type and amount of bioadhesive polymer, increase of andiroba oil, and P407 content. All formulations displayed pseudoplastic flow behavior and viscoelasticity. Bioadhesion assays in ear porcine skin showed the formulations as being equally bioadhesive. Transmission electron cryomicroscopy (Cryo-TEM) analysis showed well-structured micelles, with a size < 200 nm. The use of C974P (0.20 %, w/w) and P407 (17.5 %, w/w) constitutes a good strategy to produce emulgel systems for local administration of Andiroba oil from 4 to 8 % (w/w).

MRNA-Loaded Lipid Nanoparticles Targeting Immune Cells in the Spleen for Use as Cancer Vaccines.

mRNA delivery has recently gained substantial interest for possible use in vaccines. Recently approved mRNA vaccines are administered intramuscularly where they transfect antigen-presenting cells (APCs) near the site of administration, resulting in an immune response. The spleen contains high numbers of APCs, which are located near B and T lymphocytes. Therefore, transfecting APCs in the spleen would be expected to produce a more efficient immune response, but this is a challenging task due to the different biological barriers. Success requires the development of an efficient system that can transfect different immune cells in the spleen. In this study, we report on the development of mRNA-loaded lipid nanoparticles (LNPs) targeting immune cells in the spleen with the goal of eliciting an efficient immune response against the antigen encoded in the mRNA. The developed system is composed of mRNA loaded in LNPs whose lipid composition was optimized for maximum transfection into spleen cells. Dendritic cells, macrophages and B cells in the spleen were efficiently transfected. The optimized LNPs produced efficient dose-dependent cytotoxic T lymphocyte activities that were significantly higher than that produced after local administration. The optimized LNPs encapsulating tumor-antigen encoding mRNA showed both prophylactic and therapeutic antitumor effects in mice.

Chemoattractant releasing microneedles for enhanced DNA vaccination

Transcutaneous delivery of a DNA vaccine using microneedle (MN) has attracted much attention since there are a variety of immune cells underneath the skin. However, the clinical use of DNA vaccine is limited due to its poor cellular uptake of naked plasmid DNA (pDNA) and suboptimal presentation of an antigen by antigen presenting cells (APC). Herein, we designed a MN system for delivering polyplex-based DNA vaccine and recruiting APCs by a chemoattractant N-formyl-methionyl-leucyl-phenylalanine peptide (fMLP)-releasing microspheres (fMLP-MS) using MN coated with pH-responsive polyelectrolyte multilayer assembly (PMA). The PMA can be rapidly disrupted upon the application of MN to the skin, leading to the release of polyplex and fMLP-MS. The polyplex would be taken up by the epidermal cells and resident APCs to express and secrete the antigen (amyloid beta monomer, Aβ1-42) encoded in the DNA vaccine. fMLP released from fMLP-MS could recruit APCs to the site of MN administration by chemotactic effects. The MN-based vaccination system was able to induce robust antigen-specific antibody production.

Population pharmacokinetic/pharmacodynamic modelling of eplontersen, an antisense oligonucleotide in development for transthyretin amyloidosis.

Transthyretin-mediated amyloidosis is a progressive and fatal disease caused by the build-up of misfolded transthyretin (TTR) protein. Eplontersen is a triantennary N-acetyl galactosamine (GalNAc3)-conjugated antisense oligonucleotide targeting TTR messenger ribonucleic acid (mRNA) to inhibit production of both variant and wild-type TTR. We aimed to develop a population pharmacokinetic/pharmacodynamic (PK/PD) model for eplontersen and to evaluate the impact of covariates on exposure and response. Plasma eplontersen and serum TTR concentration data were obtained from two phase 1 studies in healthy volunteers (ClinicalTrials.gov: NCT03728634, NCT04302064). Model development was conducted using a nonlinear mixed-effects approach. Eplontersen PK was well described by a two-compartment model. Evaluation of demographics identified significant covariates of lean body mass on clearance and body weight on intercompartmental clearance and volumes of distribution. Population PK modelling showed the absorption rate was 29.6% greater with injection into the abdomen versus the arm. The typical population terminal elimination half-life was 25.5days. Serum TTR was well described by an indirect response model with inhibition of TTR production by eplontersen. Maximum fractional inhibition (Imax ) was 0.970 (0.549%RSE) and the half maximal inhibitory concentration (IC50 ) was 0.0283 ng/ml (13.3%RSE). Simulations showed subjects with lower weight had higher exposure (AUC, Cmax ), while higher Cmax was observed when comparing site of administration (ratio abdomen/arm = 1.18), but differences in exposure did not significantly impact response at evaluated doses. The exposure-response relationship of eplontersen was well characterised by the PKPD model. Weight and injection site were found to affect systemic exposure, but this effect does not seem to result in clinically relevant variation in response.

Safety and feasibility of an HPV therapeutic vaccine (TA-CIN) in patients with HPV16 associated cervical cancer (458)

<b>Objectives:</b> Therapeutic vaccines against the Human Papilloma Virus (HPV), the causative agent of cervical cancer, have the potential to stimulate HPV-specific CD8 T cell immune responses. TA-CIN is a single fusion protein comprised of HPV16 E6, E7, and L2 proteins linked in tandem. In preclinical studies, vaccination with TA-CIN induces immunity to HPV16 E6/E7-specific T cell-mediated immune responses and L2-specific neutralizing antibodies in mice. Administration of the vaccine following chemotherapy with cisplatin or radiation was able to improve the therapeutic antitumor effect in a preclinical model compared to vaccination, chemotherapy, or chemoradiation alone (Tseng et al. <i>Cancer Immunol Immunother.</i> 58:737, 2009). This suggests that a therapeutic HPV vaccine may have value in improving the therapeutic effect of the standard care of chemoradiation for patients with cervical cancer. Furthermore, preclinical studies demonstrated that site of administration in the mice influenced the extent of antitumor immunity with vaccination in the hind leg resulting in improved efficacy compared to front leg vaccination (Qiu et al. <i>Virology.</i> 525:205, 2018). This randomized, multi-center study is designed to evaluate the safety and feasibility of administering TA-CIN to patients with HPV-16 associated cervical cancer after completion of definitive primary therapy and assess the immune response when the vaccine is administered in the thigh versus the arm. <b>Methods:</b> Eligible patients for this study have HPV16-related stage IB1-IVA cervical cancer and have completed definitive treatment within the past 12 months. HPV16 nucleic acid within the cervical tumor specimen must be documented by in situ hybridization. Patients must not have evidence of disease recurrence based on prevaccination imaging and clinical assessments within eight weeks of enrollment. Other key eligibility criteria include ECOG performance status ≤1 and absolute lymphocyte count > 500/cu mm. Patients with autoimmune conditions or on immunosuppressive therapy are excluded. Fourteen patients will be randomized to receive up to 3 doses of 100ug TA-CIN vaccine three times to either the arm or the thigh. Immune and clinical responses in patients receiving vaccines will be assessed, and the injection site that elicits more potent immune responses will be selected for future studies.

Assessment of joint pharmacokinetics and consequences for the intraarticular delivery of biologics.

Intraarticular (IA) injections provide the opportunity to deliver biologics directly to their site of action for a local and efficient treatment of osteoarthritis. However, the synovial joint is a challenging site of administration since the drug is rapidly eliminated across the synovial membrane and has limited distribution into cartilage, resulting in unsatisfactory therapeutic efficacy. In order to rationally develop appropriate drug delivery systems, it is essential to thoroughly understand the unique biopharmaceutical environments and kinetics in the joint to adequately simulate them in relevant experimental models. This review presents a detailed view on articular kinetics and drug-tissue interplay of IA administered drugs and summarizes how these can be translated into reasonable formulation strategies by identification of key factors through which the joint residence time can be prolonged and specific structures can be targeted. In this way, pros and cons of the delivery approaches for biologics will be evaluated and the extent to which biorelevant models are applicable to gain mechanistic insights and ameliorate formulation design is discussed.

Reducing paclitaxel hypersensitivity reactions in gynecologic oncology patients (521)

Reducing paclitaxel hypersensitivity reactions in gynecologic oncology patients (521)

Active surveillance of adverse events after immunization (AEFI) from the Local Health Unitof Ferrara, Italy.

SummaryIntroductionVaccine vigilance implies the collection, evaluation, analysis and communication of adverse events following immunization (AEFI) and is a useful tool for vaccine monitoring allowing, even after approval and marketing, to check its safety/tolerability. The multiregional project “Active surveillance of adverse vaccine reactions”, joined by the AUSL of Ferrara, is aimed at making parents of children, who have undergone at least one vaccination provided by the regional vaccination calendar in the first 24 months of life, aware of the reporting of any AEFI via mobile phone-SMS.MethodsAn analysis of the project data, collected in the period March 2018 - May 2019, was carried out, to evaluate the effectiveness of the reporting tool and the type and frequency of AEFI. Anonymized data were analyzed by number, gender, distribution by age, type of vaccine, adverse event, severity and outcome.ResultsA total of 1,494 consents and 983 SMS messages were obtained from parents. The vaccine doses carried out were 1,984 (28.3% hexavalent, 28% PCV13, 17% anti-rotavirus, 14.3% Men-B). Almost all (99.5%) AEFI were classified as “not serious”. Based on the Organ System Class (SOC), most reports are related to “General Disorders and Administration Site Conditions” (52.3%), followed by “Psychiatric Disorders” (26.5%) and “Metabolic and nutrition disorders” (12.5%).ConclusionsThe reported AEFI are in line with the ones reported in the literature. Reporting via SMS is a valid vaccine surveillance tool contributing to the qualitative and quantitative improvement of the information transmitted.

Recent Advances in Ocular Therapy by Hydrogel Biomaterials

Current clinical practice in ocular disease treatment dosage forms primarily relies on eye drops or eye ointments, which face significant challenges in terms of low bioavailability profiles, rapid removal from the administration site, and thus ineffective therapeutic efficiency. Hydrogel has several distinct properties in semi-solid thermodynamics and viscoelasticity, as well as diverse functions and performance in biocompatibility and degradation, making it extremely promising for overcoming the challenges in current ocular treatment. In this review, the most recent developments in the use of hydrogel biomaterials in ocular therapy are presented. These sophisticated hydrogel biomaterials with diverse functions, aimed at therapeutic administration for ocular treatment, are further classified into several active domains, including drug delivery system, surface repair patch, tissue-engineered cornea, intraocular lens, and vitreous substitute. Finally, the possible strategies for future design of multifunctional hydrogels by combining materials science with biological interface are proposed.

A suicide attempt by intramuscular injection of pentobarbital sodium into rectus abdominis suggested by computed tomography.

Suicide attempts in humans due to injections of the veterinary drug pentobarbital sodium have been rarely reported. Herein, we present a case of a suicide attempt by intramuscular injection of pentobarbital sodium into the rectus abdominis muscle, which was suggested by computed tomography (CT). A 73-year-old man was brought to the emergency department with GCS 3 (E1V1M1) and an incised wound on the right side of the neck. A bottle of Somnopentyl® (pentobarbital sodium, 64.8mg/ml), a 20-ml empty syringe with an 18-mm needle, and no. 10 scalpel were present at the scene. At the emergency department, the patient was intubated and was admitted to the intensive care unit. A urine drug screen test by SIGNIFY® ER was positive for benzodiazepines and barbiturates, and continuous veno-venous hemofiltration (CHF) was initiated. The route of drug administration was initially unknown; however, a CT scan revealed swelling of the left rectus abdominis muscle with a wound suggestive of a needle puncture, and the CT analysis suggested 38.16ml as the maximum dose of pentobarbital sodium. On day 3, the patient's consciousness improved, and he was weaned off CHF and mechanical ventilation. There have been several reports of postmortem CT yielding information on the site of administration of intoxicants, but there have been none for surviving intoxicated patients. This is the first report of the usefulness of CT to identify the site of administration of the causative agent of intoxication while the patient is still alive.

Discovery of Clinical Candidate CHF-6366: A Novel Super-soft Dual Pharmacology Muscarinic Antagonist and β2 Agonist (MABA) for the Inhaled Treatment of Respiratory Diseases.

The development of molecules embedding two distinct pharmacophores acting as muscarinic antagonists and β2 agonists (MABAs) promises to be an excellent opportunity to reduce formulation issues and boost efficacy through cross-talk and allosteric interactions. Herein, we report the results of our drug discovery campaign aimed at improving the therapeutic index of a previous MABA series by exploiting the super soft-drug concept. The incorporation of a metabolic liability, stable at the site of administration but undergoing rapid systemic metabolism, to generate poorly active and quickly eliminated fragments was pursued. Our SAR studies yielded MABA 29, which demonstrated a balanced in vivo profile up to 24 h, high instability in plasma and the liver, as well as sustained exposure in the lung. In vitro safety and non-GLP toxicity studies supported the nomination of 29 (CHF-6366) as a clinical candidate, attesting to the successful development of a novel super-soft MABA compound.

580 Biodegradable bioadhesive nanoparticle delivery of chemotherapy for the treatment of cutaneous malignancies

Encapsulation of chemotherapeutic agents within biodegradable nanoparticles (NP) allows for gradual drug release from the polylactic acid core. Bioadhesive, degradable NPs can be retained at the site of administration, maximizing local effects while minimizing systemic side effects. Our bioadhesive NP (BNP) loaded with chemotherapy have previously been proven efficacious in the treatment of murine PDVC57 squamous cell carcinoma (SCC) tumors, with increased intratumoral (i.t.) retention of the active agent.

The Impact of Insulin-Induced Lipodystrophy on Glycemic Variability in Pediatric Patients with Type 1 Diabetes.

Lipodystrophy is the most common dermatological complication in patients with diabetes on insulin therapy. Despite the high frequency of lipodystrophy, there are still several difficulties in giving advice about avoidance into practice among children and adolescents with type 1 diabetes and their caregivers. This cross-sectional study aims to evaluate the prevalence of insulin-induced lipodystrophy in a cohort of pediatric patients with type 1 diabetes, to identify associated clinical factors and to assess its influence on glycemic control. Two hundred and twelve patients attending our Diabetes Center during a three-month period were enrolled. The presence of lipodystrophy was assessed by inspection and palpation procedures. Demographic and clinical data including type of treatment, frequency of rotation of insulin administration sites, and glucose metrics of the previous 30 days were assessed and statistically analyzed. Prevalence of lipohypertrophy was 44.3%. Two patients were affected by lipoatrophy (0.9%). Improper rotation of insulin administration sites and low awareness on lipodystrophy were associated to the occurrence of this skin condition (p = 0.050 and p = 0.005, respectively). When comparing patients with and without lipodystrophy, a significant difference in glycemic variability parameters was detected (p = 0.036 for coefficient of variation, p = 0.029 for standard deviation score of glucose levels). Lipodystrophy still represents a common complication in patients on insulin therapy. The present study reveals its negative impact on glycemic variability. This finding emphasizes the importance of prevention strategies to minimize the occurrence of this dermatological complication that may interfere with clinical history of the disease.

Development of approaches for studying the biodistribution of a bicistronic therapeutic plasmid construct in the mouse body

Relevance. The use of gene therapy drugs for the treatment of genetic diseases and stimulation of regeneration processes is lengthy and involves repeated injections, which may lead to increased dissemination of gene therapy constructs from the injection site and undesirable ectopic expression of growth factors encoded in them. Existing approaches to study the pharmacokinetics of a drug to assess the dissemination of a gene therapy drug from the site of administration are not applicable. Objective: to evaluate the suitability of the real-time PCR method for studying the biodistribution of a promising gene therapy drug in mice during a course of use. Methods. Male F1 CBA×C57/Black mice after nerve injury were injected with the test plasmid into the denervated tibial muscle after nerve injury, as well as after 4, 9 and 13 days at a dosage of 60 and 120 μg/mouse. After 7, 14, and 28 days, organ and tissue samples were removed, total DNA was isolated, and plasmid DNA content was assessed by real-time PCR. Results. We have shown that the studied genetic construct is able to disseminate from the injection site. We have found that the peak of dissemination for this construct in the organs and tissues of the mouse is reached 14–28 days after the end of the course application, while ectopic expression of growth factors is not observed in them. Conclusion. The proposed method is specific, highly sensitive, and linear over a wide range of concentrations. Thus, it can be recommended for studying the biodistribution of potential gene therapy drugs in the body of experimental animals as part of a preclinical studies complex.

Effect of Cyclosporine on the Body Weight of Mice Inoculated with Prostate Cancer Tissues

Cancer is a disease characterized by uncontrolled cell proliferation in the body, leading to impaired functions and high mortality. Different models are used to study cancer in vivo and in vitro. Facilities for many of the studies are not available in developing countries. Genetically modified mice for in vivo model may not be available. Cyclosporine is an immunosuppresant drug that may be used to alter mice immunity necessary for development of implanted xenograft cancer tissues. Mice were randomly separated into four groups of five and subcutaneously administered cyclosporine at different doses (5, 20 and 40 mg/kg including negative control) daily for twentyone days. At seven days of drug administration, cancer tissues from prostate cancer human patients were administered to all mice. Body weights of mice were measured at weekly interval. Tumor-like nodules were observed at the site of cancer inoculation and when present were measured. Gross pathology and histopathology were performed. The results showed significant (p&lt; 0.05) increase in body weight gain in only control group where cyclosporine was not administered. The number of tumor-like growth (first observed at day 14) per group and the magnitude of swelling vary directly (p &lt; 0.05) with cyclosporine dose. Mortality varies inversely with the dose. No lesions were observed from the gross pathology. Histopathology revealed presence of neutrophil, plasma cells and oedema with no cancer cells detected. It was concluded that cyclosporine at the dose of 20 mg/kg in mice followed by administration of prostate cancer tissue caused decrease in body weight gain and allow growth of nodule bulging at the site of cancer tissue administration..

Hyaluronic Acid-PEG-Based Diels-Alder In Situ Forming Hydrogels for Sustained Intraocular Delivery of Bevacizumab.

Retinal diseases are the leading cause of visual impairment worldwide. The effectiveness of antibodies for the treatment of retinal diseases has been demonstrated. Despite the clinical success, achieving sufficiently high concentrations of these protein therapeutics at the target tissue for an extended period is challenging. Patients suffering from macular degeneration often receive injections once per month. Therefore, there is a growing need for suitable systems that can help reduce the number of injections and adverse effects while improving patient complacency. This study systematically characterized degradable “in situ” forming hydrogels that can be easily injected into the vitreous cavity using a small needle (29G). After intravitreal injection, the formulation is designed to undergo a sol–gel phase transition at the administration site to obtain an intraocular depot system for long-term sustained release of bioactives. A Diels–Alder reaction was exploited to crosslink hyaluronic acid-bearing furan groups (HAFU) with 4 arm-PEG10K-maleimide (4APM), yielding stable hydrogels. Here, a systematic investigation of the effects of polymer composition and the ratio between functional groups on the physicochemical properties of hydrogels was performed to select the most suitable formulation for protein delivery. Rheological analysis showed rapid hydrogel formation, with the fastest gel formation within 5 min after mixing the hydrogel precursors. In this study, the mechanical properties of an ex vivo intravitreally formed hydrogel were investigated and compared to the in vitro fabricated samples. Swelling and degradation studies showed that the hydrogels are biodegradable by the retro-Diels–Alder reaction under physiological conditions. The 4APM-HAFU (ratio 1:5) hydrogel formulation showed sustained release of bevacizumab > 400 days by a combination of diffusion, swelling, and degradation. A bioassay showed that the released bevacizumab remained bioactive. The hydrogel platform described in this study offers high potential for the sustained release of therapeutic antibodies to treat ocular diseases.